OBJECTIVE: This study examined the expression of insulin-like growth factor-1(IGF-1)and its receptor in 3-day-old premature rats with chronic hypoxic ischemic brain damage (HIBD) and investigated the role of IGF-1 in the pathogenesis of this disease. METHODS: Ninety 3-day-old Sprague-Dawley rats were randomly assigned into a Control group (n =40) and a Hypoxic-ischemic group (HI, n = 50). HI was induced through bilateral common carotid artery ligation. The Control group was only sham-operated. Hematoxylin and eosin staining, TUNEL immunofluorescence staining and immunohistochemistry ways were used to investigate the expression of IGF-1 and its receptor, morphological changes of brain tissues and cell apoptosis of brain white matter. RESULTS: The expression of IGF-1 decreased in 3-5 days after HI, but that of its receptor increased in the HI group. The expression changes were most significant at corpus callosum and peri-ventricular white matter and recovered progressively in 7-14 days after HI. After 7 days of HI, the brain white matter presented with morphological changes such as rarefaction, liquefaction and lateral ventricular enlargement. Apoptotic cells in deep white matter increased after HI, and peaked at 48 hrs. CONCLUSIONS: IGF-1 may play an important role in the pathogenesis of chronic HIBD in 3-day-old premature rats. This study provides an experimental basis for the prevention and treatment of HIBD in premature infants.

OBJECTIVE: Recent studies have suggested that acute lung injury (ALI ) is an important cause of pulmonary hemorrhage of the newborn (PHN) and that surfactant protein A (SP-A) and tumor necrosis factor-α (TNF-α) are involved in the development of ALI. This study examined the levels of SPA and TNF-α in bronchoalveolar lavage fluid (BALF) and their relationship in newborns with pulmonary hemorrhage(PH). METHODS: Twenty newborn infants with PH and 15 sick neonates but without PH (Control group) were enrolled in this study. According to the prognosis, the PH group was subdivided into Survival group (n=14) and Death group (n=6). The Western-dot blot analysis and enzyme-linked immunoadsorbent assay (ELISA) were used to detect the levels of SP-A and TNF-α in BALF and serum TNF-α level. RESULTS: The SP-A levels in BALF in the survival PH patients in the acute stage and in the death PH cases were significantly lower than those of the Control group. The death PH patients showed a much lower SP-A level in BALF than the survival ones in the acute stage. Whilst the recovery stage of PH, the SP-A level in BALF in the survival patients increased significantly and remained the similar level as the Control group. The TNF-α levels in both serum and BALF in the survival PH patients in the acute stage and in the death PH cases were significantly higher than those in the Control group. The death PH cases showed a higher level of TNF-α in serum and BALF than PH survival cases in the acute stage. The increased extent of TNF-α level in BALF was obviously greater than of that in serum. In the recovery stage, the level of TNF-α in BALF was significantly reduced and almost returned to the level of the Control group in the survival PH cases compared with that in the acute stage. There was a negative correlation between the BALF SP-A and TNF-α levels in newborns with PH (r = - 0.635,P = 0.003). CONCLUSIONS: SP-A and TNF-α may be involved in the process of lung injury in PHN. Monitoring the levels of SP-A and TNF-α is useful for the early diagnosis and treatment and the evaluation of the outcome of PHN.

OBJECTIVE: To evaluate two small interfering RNAs (siRNAs) on the NPM-ALK fusion gene expression in anaplastic large-cell lymphoma cell line Karpas299, and to study the effect of RNA interference on Karpas299 cells proliferation. METHODS: Two siRNAs sequences (siRNA- I and siRNA-II) were designed to target the NPM-ALK fusion site in anaplastic large-cell lymphoma cell line Karpas299. An siRNA U6 expression system including U6 RNA-based polymerase III promoter was set up. The two siRNAs designed for down-regulation of the NPM-ALK fusion mRNA were transfected into Karpas299 cells by liposomal transfection reagents. The effect of RNAi on NPM-ALK mRNA expression was detected by real-time RT-PCR and Western blot. The anti-proliferative effects of the siRNA U6 system were assessed using MTT. Apoptosis was observed by fluorescence microscopy. RESULTS: The mRNA level of NPM-ALK in Karpas299 cells transfected with siRNA-I and siRNA-II decreased by approximately 75% and 35% respectively. The NPM-ALK protein expression was inhibited in Karpas299 cells at 72 hrs of siRNA-I transfection. The siRNA-II treatment had no effect on NPM-ALK protein expression. siRNA-I had inhibitory effects on Karpas299 cells proliferation and induced the cells apoptosis, while siRNA-II did not. CONCLUSIONS: Sequence specific siRNAs targeting NPM-ALK was capable of suppressing NPM-ALK expression and inhibiting cellular proliferation. RNA interference may be a suitable technique for studying the function of NPM-ALK gene and may be used to develop siRNA-based targeted gene therapeutic approaches against NPM-ALK-positive lymphomas.

OBJECTIVE: To investigate the dynamic changes of transforming growth factor-β_1 (TGF-β_1) mRNA and protein expression and their effects on lung development in premature rats with chronic lung disease (CLD). METHODS: CLD was induced by hyperoxia exposure (FiO_2=0.90) in neonatal premature rats. The dynamic changes of lung coefficient and radical alveolar counts (RAC) were observed; and the expressions of TGF-β_1 mRNA and protein were assayed with reverse transcription polymerase chain reaction (RT-PCR), immunohistochemical and image processing technique on days 1, 3, 7, 14 and 21 after birth. For comparison analysis, 30 premature rats exposed to air were used as the Control group. RESULTS: There were no differences in the lung coefficient and the RAC between the CLD and the Control groups within 3 days after birth. However on the 7th and 14th days, the lung coefficient and the RAC of the CLD group were significantly lower than those of the Control group. The RAC decreased to the nadir on the 21st day, but the lung coefficient was not different from the Control group. In the CLD group, the expression level of TGF-β_1 mRNA increased on the 3rd day, peaked on the 14th day and remained higher on the 21st day; the higher expression of TGF-β_1 protein was detected on the 7th day and peaked on the 21st day; and the TGF-β_1 protein expression was negatively correlated with RAC(P =0.0027). CONCLUSIONS: The over-expression of TGF-β_1 in lung tissues is associated with the lung development disorder. The TGF-β_1 expression level may be an index for the assessment of lung development.

This paper reported a 13-year-old girl with hereditary megaduodenum.This disease rarely occurs in children. It is a rare familial disease with autosomal dominant transmission characterized by hollow visceral myopathy.It usually presents as recurrent intestinal pseudo-obstruction, clinically manifesting intermittent vomiting and abdominal distention,which can lead to malnutrition and growth retardation. Barium meal examination displays duodenum distention and esophageal manometry reveals esophageal dystonia. The histological features include inflammatory infiltration, marked thinning and fibrosis of the smooth muscle in the intestinal specimens. Diagnosis of the disease depends on the familial history, histological examination, radiographic and esophageal function examinations. Treatment is based on diet and control of bacterial overgrowth. Surgical operation can relieve obstructive symptoms.

OBJECTIVE: This study aimed to investigate the effect of clearance of tumor cells in autologous bone marrow after induction chemotherapy (based on RT-PCR detection) on the outcome in patients with neuroblastoma. METHODS: Thirty-two neuroblastoma patients with histological bone marrow metastasis at diagnosis but without tumor cells in marrow based on the immunohistochemistry detection after induction therapy were eligible to this study. The neuroendocrine protein gene product 9.5 (PGP9.5) was measured with RT-PCR, as a bio-marker to detect the presence of tumor cells in bone marrow at diagnosis and after induction chemotherapy. RESULTS: All patients were PGP9.5 positive in bone marrow at diagnosis. After induction chemotherapy, 16 patients remained PGP9.5 positive in marrows. A 3.4±0.9 year-follow-up showed a relapse in 11 out of the 16 patients, with event free survival (EFS) of 31%. In contrast, among the other 16 PGP9.5 negative patients, only 5 relapsed after (3.2±0.7 years) treatment, with EFS of 69%(P = 0.018). Conclusions Clearance of neuroblastoma cells through induction chemotherapy (based on RT-PCR detection) in autologous bone marrow is associated with better prognosis in patients with neuroblastoma.

OBJECTIVE: The objective of this study was to assess the prognostic value of response to prednisone in children with acute lymphoblastic leukemia (ALL). METHODS: One hundred and twenty-six children with newly diagnosed ALL were enrolled in the protocol of ALL-XH-90. A prednisone good response (PGR) was defined as a blast count of less than 1 000/μL and a prednisone poor response (PPR) as a blast count of at least 1 000/μL in peripheral smears, after 7 days of oral prednisone (60 mg/m~2) and one intrathecal injection of methotrexate along with cytosine arabinoside. The probability of event-free survival (pEFS) was estimated by Kaplan-Meier analysis and the differences in pEFS between groups were assessed by the log-rank test. Differences in the biologic characteristics between groups were compared by Chi-square analysis or Fisher exact test (two-tail). RESULTS: One hundred and ten patients had PGR and 16 had PPR. The 5-year pEFS was significantly worse for patients with PPR compared with those with PGR (48%±13% vs 73%±5%, P= 0.0021). CONCLUSIONS: Response to prednisone is a simple, practical and prognostic factor in childhood ALL.

OBJECTIVE: To evaluate the clinical significance of minimal residual disease (MRD) in childhood B-lineage acute lymphoblastic leukemia (B-ALL). METHODS: MRD was detected using triple-color flow cytometry (FCM) with a combination of specific and sensitive marker of cell plasma and membrance with CD45/SSC in 67 children with B-ALL at the end of the remission induction therapy. RESULTS: Of the 67 patients, 18 were low risk; 35, medium risk and 14 high risk. The MRD positive rate was significantly different among these patients with different degrees of risk. The high risk patients had the highest MRD positive rate, followed by the medium risk patients (P<0.05). MRD was not significantly correlated with the gender, age and the initial leukocyte counts, as well as with the response to early therapy. A significant association between MRD and the incidence of relapse as well as the event-free survival (EFS) period was observed. The relapse rate in patients with MRD positive was remarkably higher than that in those with MRD negative. The patients with MRD positive had a shorter EFS period compared with MRD negative patients. CONCLUSIONS: The detection of MRD can be used to evaluate the therapeutic effect and the prognosis of childhood ALL.

OBJECTIVE: CD40 and CD40 ligand (CD40L) are a pair of co-stimulatory molecules in immunoreaction, and might play an important role in the development of asthma. This study aimed to explore the relationship between the expression of CD40 and CD40L and the development of asthma in children. METHODS: Thirty-two children with acute asthma were selected randomly and 20 age-matched healthy children were enrolled as controls. Flow cytometry was used to detect the expression of CD40 and CD40L in T and B lymphocytes in peripheral blood. RESULTS: The expression level of CD40 in B cells of asthmatic patients was significantly higher than that of controls (17.05±4.88 vs 13.89±3.67, P < 0.05). CD4~+T cells of asthmatic patients expressed a higher level of CD40L compared with controls (8.04±3.53 vs 5.58±3.00,P <0.05) . CONCLUSIONS: Expression of CD40 and CD40L were increased in relevant immune cells of children with acute asthma, which may contribute to the immune pathogenesis of asthma.

OBJECTIVE: Single strand conformation polymorphism (SSCP) and restriction fragment length polymorphism (RFLP) have been used for the diagnosis of spinal muscular atrophy (SMA), but the two methods were complex. In order to find out a simpler and reliable method, this paper studied the feasibility of allele-specific PCR (AS-PCR) in the gene diagnosis of SMA. METHODS: AS-PCR technique was used to detect the deletion of exon 7 of survival motor neuron (SMN) gene in 40 patients with SMA (15 cases of type I, 17 type II and 8 type III) and in 40 healthy controls. All the patients were confirmed to have deletion of exon 7 of SMN_1 by the RFLP method. RESULTS: AS-PCR showed that all the 40 patients had deletion of exon 7 of SMN_1. This result was consistent with that detected by the RFLP method. CONCLUSIONS: AS-PCR technique is simple and reliable for the gene diagnosis of SMA.

OBJECTIVE: Lennox-Gastaut syndrome (LGS) is a refractory epilepsy in children. Some are accompanied by electrical status epilepticus during sleep (ESES)rendering treatment more difficult. This study examined the short-term effect of intravenous methylprednisolone (MP) pulse therapy on LGS with ESES. METHODS: Twenty-two children with LGS, who were confirmed with ESES by electroencephalograph (EEG), received MP intravenous pulse therapy followed by prednisone administration. RESULTS: Fifteen patients had partial remission of seizures (68.2%)and the EEG showed reduced epileptiform discharges in 16 cases (72.7%). No severe adverse effects were found. CONCLUSIONS: MP pulse therapy can alleviate the symptoms and ESES in children with LGS. If anti-epileptic drug treatment is not effective, the therapy should be considered.

A case of valproic acid (VPA)-induced hepatic failure was reported. An 8.5-month-old female developed infantile spasm at 1 month of life. She was then treated with high-dose VPA and clonazepam (CNP). She manifested with frequent seizures in the presence of pneumonia. Gingival bleeding, as well as multiple punctuated hemorrhagic spots were observed on her face and cervico-thoracic areas. There was severe edema on her extremities, especially the lower extremities. A significant reduction of full blood count and severe hypoproteinemia were noted although transaminase ALT was still within normal range. Both serum VPA and CNP levels were beyond the reference range. Diagnosis of VPA-induced hepatic failure was based on the clinical and laboratory findings. This study reviewed the high risk factors, the pathogenesis, early diagnosis, prevention and treatment of VPA-induced hepatic failure based on the case report and published literatures.

OBJECTIVE: Precordial distress is a common complaint in the pediatric population resulting from cardiac or non-cardiac diseases. The management approach and the prognosis of precordial distress vary with different causes. Therefore it is important to identify the causes of this complaint. This study examined serum cardiac troponin I (cTnI) levels and other clinical and laboratory parameters in children with this complaint to explore the value of cTnI in identifying the causes of childhood precordial distress. METHODS: The medical documents of 123 children with precordial distress, including the history, the findings of physical examination, electrocardiogram, ultrasonocardiography and myocardial perfusion imaging (MPI), serum creatine kinase MB isoenzyme (CK-MB) activity and cTnI concentrations, were analyzed retrospectively. cTnI concentrations were detected using enzyme immoassy. The relationship between myocardial perfusion and cTnI concentrations was evaluated by rank correlation analysis. RESULTS: A total of 81 patients (65.9%) were diagnosed with non-cardiac disease among 123 cases with precordial distress and all of them had normal cTnI concentrations. The cTnI concentrations were increased above normal in 17 cases, and low myocardial perfusion was found in 36 of 42 children with cardiac diseases. There was a linear correlation between cTnI levels and the severity of myocardial perfusion decrease (r=0.974, P<0.01). CONCLUSIONS: The majority of childhood precordial distress may be caused by non-cardiac disease. Measurement of cTnI combined with MPI can identify the cause of childhood precordial distress.

OBJECTIVE: To study the serum leptin level and its relation to physical development in children, aged between 3 and 9 years, from an undeveloped region of China. METHODS: A total of 280 children aged between 3-9 years from a kindergarten and an elementary school of an undeveloped county of Hubei Province (147 boys and 133 girls) were enrolled in this study. They were divided into four groups according the age and gender: 3-6 year-old boy / girl group and 6-9 year-old boy / girl group. Their weights and heights were measured and body mass index (BMI) and body fat precentage(BFP) were calculated. The serum leptin level was detected using ELISA and was compared between the groups. The correlation between the serum leptin level and physical development parameters was evaluated. RESULTS: Serum leptin levels in the girls were significantly higher than those in the boys in the same age group (P <0.05), but no differences were found between children aged 3-9 years of the same gender. The leptin level was obviously correlated with BMI and BFP in both boys and girls aged 6 to 9 years (P<0.05), but not in children aged 3 to 6 years. CONCLUSIONS: The serum leptin level was different between boys and girls in children aged 3 to 9 years in the undeveloped region. It was correlated to BMI and BFP in children aged 6 to 9 years.

Objective To evaluate the levels and clinical significance of serum epidermal growth (EGF) in premature infants.Methods Serum EGF concentrations of 10 full-term and 35 premature infants were measured by radio-immunoassay (RIA ) within 24hrs after birth.Results The serum EGF concentrations in premature infants born between 35 and 37 weeks of gestation(0.617±0.22 μg/L)and those born between 28 and 34 weeks of gestation(0.540±0.31 μg/L)were significantly lower than those in full-term infants(0.723±0.18 μg/L)( P<0.01). The asphyxiated premature infants showed a much lower EGF level(0.446±0.24 μg/L) compared with the full-term infants. The EGF concentration of the premature infants with necrotizing enterocolitis was not significantly different from that of the full-term infants. Conclusions The serum EGF concentration is associated with the gestational age. Asphyxia may result in a decreased EGF level.

OBJECTIVE: The neonatal rat model of hypoxic-ischemic brain damage (HIBD) is a useful tool for studying hypoxic-ischemic encephalopathy of the newborn. Although many reports have studied the pathological and biochemical outcomes on HIBD in the animal model, little research has focused on assessing the long-lasting behavioral changes. This study was designed to examine the long-lasting behavioral alternations following HIBD in neonatal rats. METHODS: Twenty-four 7-day-old SD rats were randomly assigned into a Control group (n = 12) and a HIBD group (n = 12). The rats in the HIBD group were subjected to ligation of the left carotid artery, followed by 2 hrs hypoxia exposure. A battery of behavioral tests, including the T-maze test and sensorimotor tests, were performed at postnatal age 3 - 4 weeks. Histological changes and their correlations with the results of behavioral tests were evaluated. RESULTS: In the behavioral tests, HIBD rats performed significantly worse than control rats. In the T-maze test, the HIBD group differed significantly from the Control group, achieving the correct percentage on the 3rd and 4th days after hypoxia-ischemia (HI) of 68.3%±26.2% vs 86.7%±15.6% ( P =0.049) and 66.7%±15.6% vs 98.3%±5.7% (P < 0.001), respectively. In the sensorimotor tests, asymmetries of foot-faults and limb-placing were detected in HIBD rats. The postural reflex test showed an abnormal motor function in HIBD rats. The neurons number in the DG area of the hippocampus (39.7 ± 5.9) and the cortex ( 12.7±3.3) was significantly reduced in the HIBD group when compared with those in the Control group (50.9 ± 4.1 and 18.2 ± 3.3, respectively). There was no correlation between the behavioral tests results and histological changes. CONCLUSIONS: HI injury in neonatal rats may cause long-lasting deficits of learning, spatial and sensorimotor function. Behavioral testing is an important measure of outcome following HIBD, and behavioral testing measures may be used to test the efficacy of HIBD treatment.

OBJECTIVE: This study was aimed to investigate the expressions of ICAM-1 and TNF-α in viral myocarditis and the effect of anti-TNF-alpha monoclonal antibody on viral myocarditis in mice. METHODS: Sixty-six male Balb/c mice were randomly assigned into an Infection group (n=30), an Intervention group (n=18) and a Normal control group (n=18). The Infection and Intervention groups were inoculated intraperitoneally with 0.2 mL TCID50 10~ -6/mL coxsackie B_3(CVB_3) solution. Anti-TNF-amAb [0.1 mL/(kg.d)] was additionally administered starting at 1 day before CVB_3 virus inoculation until day 5 in the Intervention group. On the 7th and 14th days after virus inoculation, the changes of histopathology and ultrastructure of myocardium were studied with light and electron microscopy. The expressions of ICAM-1 and TNF-α were detected by immunohistochemical method. RESULTS: Myocardium histopathology of mice in the Normal control group was normal. Myocardial necrosis and cellular infiltration were more prominent in the Infection group than in the Intervention group.ICAM-1 and TNF-α were expressed in the myocardium of Normal control gorup at a very low level, which were significantly lower than those in the Infection and Intervention groups. The expressions of ICAM-1 and TNF-α were dramatically reduced in the Intervention group compared with those in the Infection group.In the Infection group, a positive correlation was found between the expressions of ICAM-1 and TNF-α ( r=0.706, P<0.05); and both ICAM-1 expression and TNF-α expression were positively related to pathological scores of myocardium (r = 0.737, P<0.05;r = 0.693, P<0.05). CONCLUSIONS: ICAM-1 and TNF-α may play important roles in the pathogenesis of viral myocarditis. Anti-TNF-alpha monoclonal antibody has protective effects on myocardial tissues by inhibiting the ICAM-1 and TNF-α expression.

OBJECTIVE: The effects of nuclear factor-kappaB (NF-κB) on neurons in the brain remain controversial. This study examined the relationship between NF-κB activation and neuronal apoptosis in rats with pentylenetetrazol (PTZ)-induced serious seizures. METHODS: Forty-two rats were randomly assigned into a Seizure group, a pyrrolidine dithiocarbamate (PDTC)-pretreatment seizure group and a Normal control group. The model of serious seizures was induced by an intraperitoneal injection of PTZ (60 mg/kg). Rats in the Seizure group were sacrificed 3, 6, 12, 24, and 48 hrs after seizure induction. The PDTC pretreatment group was administered with PDTC (100 mg/kg) 1 hr before PTZ injection, and sacrificed 24 hrs later. The immunohistochemistry method was used to detect the activity of NF-κB protein in the CA1 region of the hippocampus. The in situ dUTP end-labeling (TUNEL) technique and flow cytometry (FCM) were used to measure the neuronal apoptosis in the hippocampus. RESULTS: Significant neuronal apoptosis was observed in the Seizure group and peaked at 24 hrs, with an apoptosis rate of 12.54%±4.99%, compared with (2.24%±0.57%) in the Normal control group (P<0.001). The activity of NF-κB was detected at 6 hrs following seizure induction, and increased to maximal expression at 24 hrs (32.30±4.71/gcs)in the Seizure group. NF-κB activation was not found in the Normal control group. The PDTC pretreatment group showed a lower neuronal apoptosis rate and less NF-κB activity than the Seizure group. CONCLUSIONS: The activation of NF-κB may play important roles in the process of neuronal apoptosis following serious seizures. PDTC pretreatment could reduce the neuronal apoptosis, perhaps by inhibiting the expression of NF-κB.

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